[The use of atomic-force microscopy for the analysis of microbiological objects].
Atomic force microscopy (AFM) was used for the analysis of rickettsiae and viruses. The specificity of interaction was evaluated on the basis of the adsorption of the analyzed antigen on the polymer-antibody film. The film was formed and transferred onto highly oriented pyrolytic graphite (HOPG) by the method of Langmuir-Schaefer with the use of amphiphilic polymers, alkylated polyethyleneimines. According to the data of AFM, polymer-antibody film was 10-30 nm thick and had ruptures, uneven surface. AFM images of Coxiella burnetii, rotavirus and Venezuelan equine encephalitis virus, immunoadsorbed on the antibody film, were obtained. C.burnetii, due to their size equal to (700-900) x (300-500) x Z = 200 nm, were clearly visible on the underlying surface and could be directly counted. Individual virus particles (60-80 nm) cold not be identified on such surface. To analyze such preparations, the program of image analysis was developed. The program classified the registered image with a certain standard. This program determined the presence of virus antigen on the underlying affinity surface with a high degree of precision.
Budashov
I A
IA
All-Russian Center of Molecular Diagnostics and Treatment, Moscow, Russia.
Kurochkin
I N
IN
Denisov
A K
AK
Skripniuk
V V
VV
Shabanov
T A
TA
D000327:Adsorption; D016997:Coxiella burnetii / Q000648:ultrastructure; D004666:Encephalitis Virus, Venezuelan Equine / Q000648:ultrastructure; D008828:Microbiological Techniques / Q000295:instrumentation; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D012401:Rotavirus / Q000648:ultrastructure; D014616:Vaccinia virus / Q000648:ultrastructure
Ispol'zovanie atomno-silovoĭ mikroskopii dlia analiza mikrobiologicheskikh ob''ektov.